Generation of redesigned homing endonucleases comprising DNA-binding domains derived from two different scaffolds.

ABSTRACT: Homing endonucleases have become valuable tools for genome engineering. Their sequence recognition repertoires can be expanded by modifying their specificities or by creating chimeric proteins through domain swapping between two subdomains of different homing endonucleases. Here, we show that these two approaches can be combined to create engineered meganucleases with new specificities. We demonstrate the modularity of the chimeric DmoCre meganuclease previously described, by successfully assembling mutants with locally altered specificities affecting both I-DmoI and I-CreI subdomains in order to create active meganucleases with altered specificities. Moreover these new engineered DmoCre variants appear highly specific and present a low toxicity level, similar to I-SceI, and can induce eff

SUBMITTER: Grizot S 

PROVIDER: S-EPMC2847234 | biostudies-literature | 2010 Apr

REPOSITORIES: biostudies-literature